| Project/Area Number |
14370353
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General surgery
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| Research Institution | Kyoto University |
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Principal Investigator |
IKAI Iwao Kyoto University, Department of Gastroenterological Surgery, Lecturer, 医学研究科, 講師 (60263084)
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| Co-Investigator(Kenkyū-buntansha) |
HIROSE Tetsuro Kyoto University, Department of Gastroenterological surgery, Assistant, 再生医科学研究所, 助手 (00314279)
MITAKA Toshihiro Sapporo Medical University School of Medicine, Department of Pathophysiology, Cancer Research Institute, Professor, がん研究所, 教授 (50231618)
YAMAOKA Yoshio Kyoto University, Department of Gastroenterological surgery, Professor, 医学研究科, 教授 (90089102)
UESUGI Takehiko Kyoto University, Department of Gastroenterological surgery, Assistant, 医学研究科, 助手 (20362494)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥14,700,000 (Direct Cost: ¥14,700,000)
Fiscal Year 2003: ¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 2002: ¥7,300,000 (Direct Cost: ¥7,300,000)
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| Keywords | hepatic stem cell / human liver / tissue reconstruction / hepatic progenitor cell / non-parenchymal cell / mesenchymal cell / ヒト小型肝細胞 / 1型コラーゲンシート / 共培養 / 類肝組織構築 / コラーゲンタイプIシート |
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| Research Abstract |
The aim of this study was to isolate and characterize human hepatic stem/progenitor cells from human liver and to reconstruct the liver tissue.
Isolation method of hepatic stem progenitor cells from fetal and adult mise liver was established using cell aggregation method: We characterized three populations in the cell aggregations from the liver of fetal mice using specific cell surface markers Thy-1 positive mesenchymal cells; CD49f positive hepatic progenitor cells and CD45-positive blood cells, Thy-1 positive cells promoted (differentiation of C D49f positive cells to mature hepatocytes. Hepatic progenitor cells from the liver of green fluorescent protein (GFP) transgenic adult mice were also isolated by fluorescent activated cell sorting. We select 30 genes. for candidates of surface markers of hepatic stem/progenitor cells using cDNA subtraction between GFP positive hepatic progenitor cells and normal mature hepatocyte. Three of 30 genes were expressed in the fetal mouse hepatic progenitor cells specifically. Bone marrow cells in mice, which have multi-lineage differentiation activity, were committed to hepatic sinusoidal endothelial cell and Kupffer cells during liver regeneration and to hepatic stellate cells during liver fibrosis.
We developed a suitable method for human hepatocytes isolated from resected specimens to maintain both proliferative capacity and differentiated functions. Three-dimensional hepatic organoid was reconstructed using co-culture of hepatocytes and hepatic non-parenchymal cells isolated from resected specimens on collagen type I sheet.
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